Terni Beatrice, Pacciolla Paolo, Perelló Margalida, Llobet Artur
Laboratory of Neurobiology, Neuroscience Program, Bellvitge Biomedical Research Institute (IDIBELL), L'Hospitalet de Llobregat; Department of Pathology and Experimental Therapy, School of Medicine, University of Barcelona, L'Hospitalet de Llobregat; Institute of Neurosciences, University of Barcelona, L'Hospitalet de Llobregat.
Laboratory of Neurobiology, Neuroscience Program, Bellvitge Biomedical Research Institute (IDIBELL), L'Hospitalet de Llobregat; Department of Pathology and Experimental Therapy, School of Medicine, University of Barcelona, L'Hospitalet de Llobregat; Institute of Neurosciences, University of Barcelona, L'Hospitalet de Llobregat;
J Vis Exp. 2018 Dec 11(142). doi: 10.3791/58028.
Xenopus tadpoles offer a unique platform to investigate the function of the nervous system. They provide multiple experimental advantages, such as accessibility to numerous imaging approaches, electrophysiological techniques and behavioral assays. The Xenopus tadpole olfactory system is particularly well suited to investigate the function of synapses established during normal development or reformed after injury. Here, we describe methodologies to evaluate the processing of olfactory information in living Xenopus larvae. We outline a combination of in vivo measurements of presynaptic calcium responses in glomeruli of the olfactory bulb with olfactory-guided behavior assays. Methods can be combined with the transection of olfactory nerves to study the rewiring of synaptic connectivity. Experiments are presented using both wild-type and genetically modified animals expressing GFP reporters in central nervous system cells. Application of the approaches described to genetically modified tadpoles can be useful for unraveling the molecular bases that define vertebrate behavior.
非洲爪蟾蝌蚪为研究神经系统功能提供了一个独特的平台。它们具有多种实验优势，例如可采用多种成像方法、电生理技术和行为分析方法。非洲爪蟾蝌蚪的嗅觉系统特别适合研究正常发育过程中建立的或损伤后重新形成的突触的功能。在这里，我们描述了评估活体非洲爪蟾幼体嗅觉信息处理的方法。我们概述了在嗅球肾小球中对突触前钙反应进行体内测量与嗅觉引导行为分析相结合的方法。这些方法可与嗅神经横断相结合，以研究突触连接的重新布线。我们展示了使用野生型和在中枢神经系统细胞中表达绿色荧光蛋白报告基因的转基因动物进行的实验。将所描述的方法应用于转基因蝌蚪，可能有助于揭示定义脊椎动物行为的分子基础。
